Sutoh K
Department of Pure and Applied Sciences, College of Arts and Sciences, University of Tokyo, Japan.
Plasmid. 1993 Sep;30(2):150-4. doi: 10.1006/plas.1993.1042.
A new selectable marker for transformation of Dictyostelium discoideum cells was constructed by using the bsr gene from Bacillus cereus, which confers resistance to Blasticidin S. The bsr gene was driven by Dictyostelium actin 15 promoter and Dictyostelium actin 8 terminator for expression in Dictyostelium cells. To demonstrate the feasibility of using the bsr marker, we constructed an extrachromosomal replication vector by replacing the Neor gene of pnDeI (B. Leiting and A. Noegel (1988) Plasmid 20, 241-248) with the bsr gene cassette. A mutant Dictyostelium actin 15 gene was constructed and inserted into the vector. Dictyostelium cells were transformed with the resulting vector and then transformants were selected with Blasticidin S. The selected cells showed high level expression of the mutant actin, indicating an efficient selection of transformed cells with the bsr marker.
通过使用蜡状芽孢杆菌的bsr基因构建了一种用于盘基网柄菌细胞转化的新选择标记，该基因赋予对杀稻瘟菌素S的抗性。bsr基因由盘基网柄菌肌动蛋白15启动子和盘基网柄菌肌动蛋白8终止子驱动，以便在盘基网柄菌细胞中表达。为了证明使用bsr标记的可行性，我们通过用bsr基因盒替换pnDeI（B. Leiting和A. Noegel（1988年）《质粒》20卷，241 - 248页）的Neor基因构建了一个染色体外复制载体。构建了一个突变的盘基网柄菌肌动蛋白15基因并插入到该载体中。用所得载体转化盘基网柄菌细胞，然后用杀稻瘟菌素S选择转化体。所选细胞显示出突变肌动蛋白的高水平表达，表明用bsr标记能有效选择转化细胞。
